Simultaneous Determination of Six Isoflavonoids in Radix Puerariae Lobatae from Different Habitats by HPLC

Objective:A reversed-phase high performance liquid chromatographic method(RP-HPLC) was developed for the simultaneous analysis of six isoflavonoids of Gegen,the roots of Puerariae lobatae.Method:HPLC determination was performed on a Kromasil C18 column(4.6 mm×250 mm,5 μm) and detected at 250 nm.The mobile phase was consisted of methanol and 0.1% citric acid solution with gradient elution.The flow rate was 1.0 mL·min-1,and column temperature was kept at 30 ℃.Result:The method was proved to be linear in the ranges of 76.2-1 016,65.8-1 316,85.6-1 712,49.98-499.8,21.94-219.4,48.5-485 ng for the six isoflavonoid,3′-hydroxypuerarin,puerarin,daidzin,genistin,ononin,daidzein,respectively.The average recoveries were 99.73%(RSD 0.6%),100.48%(RSD 1.11%),100.37%(RSD 0.79%),100.28%(RSD 1.07%),99.96%(RSD 1.22%),97.36%(RSD 1.59%),respectively.Conclusion:Results obtained showed that the method is convenient,accurate and reliable,can be applied to the simultaneous determination of the six isoflavonoids in Radix Puerariae lobatae.